Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
  • A61K39/39591—Stabilisation, fragmentation
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L2/00—Disinfection or sterilisation of materials or objects, in general; Accessories therefor
  • A61L2/02—Disinfection or sterilisation of materials or objects, in general; Accessories therefor using physical processes
  • A61L2/04—Heat
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L2103/00—Materials or objects being the target of disinfection or sterilisation
  • A61L2103/05—Living organisms or biological materials
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
  • Y10S530/861—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof involving IgG3, IgG4, IgA, or IgY
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
  • Y10S530/863—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof involving IgM

Definitions

• This invention relates to a process for heat-treating immunoglobulin to inactivate the virus possibly present therein.
• liquid heating method The most reliable method heretofore used for treating plasma protein such as albumin to inactivate the virus which possibly contaminates the protein is to heat-treat it in the state of its aqueous solution (hereinafter referred to as "liquid heating method"), which is based on the report of Murray et al. [The New York Academy of Medicine, 31 (5), 341-358 (1955)]. The method has been widely used through many years up to the present and the effect of inactivating viruses of the liquid heating method has been substantiated also epidemilogically.
• albumin of plasma proteins can withstand the liquid heating.
• those plasma proteins which have physiological or biological activities are very sensitive to heat, susceptible to thermal denaturation, and liable to undergo the decrease or disappearance of the activities.
• the stabilizer such as albumin, glycine, mannitol or sorbitol does not affect on the stability of the blood coagulation enzyme but gives good solubility and clarity to an aqueous solution of the virus-inactivated enzyme after the heat treatment.
• An object of this ihvention is to provide a dry heat treatment of immunoglobulin, which can inactivate viruses present therein without inactivating the immunoglobulin itself.
• the present inventors have found that viruses present in immunoglobulin can be inactivated without losing the activity of the globulin by dry-heat treating the globulin and that when the dry heat treatment of immunoglobulin is conducted in the presence of a stabilizer the globulin is markedly stabilized further and the immunoglobulin dry-heat treated under such conditions shows good solubility in water and good state of solution.
• This invention has been accomplished on the basis of these findings.
• this invention relates to a process for heat-treating immunoglobulin which comprises heating immunoglobulin contaminated with a virus in a substantially dry state, preferably in the presence of a specified stabilizer, until the virus becomes inactivated, whereby the contaminant virus is inactivated and the solubility in water and the stability of the immunoglobulin are improved.
• immunoglobulins which are the subject of the heat treatment of this invention are those which have biological or physiological activities characteristic of immunoglobulin, for example, those which can be obtained by the fractionation of plasma proteins.
• immunoglobulins examples include those of human, horse and mouse origin. These may be either a polyclonal antibody or a monoclonal antibody and are preferably IgG, IgA or IgM.
• This invention is usually carried out by lyophilizing an immunoglobulin solution and then heating the lyophilized product in substantially dry state, that is, a state with as small moisture content as possible, preferably at 3% or less, and usually 0.05 to 3%.
• the temperature used in the present dry heat treatment is usually 30° to 100° C., most preferably about 60°, and the heating time is one sufficient for inactivating the virus only, and it is usually 10 minutes to 200 hours depending on the temperature used, and preferably about 10 to 100 hours.
• viruses which are to be the object of the inactivation by heat treatment of this invention are those which are suspected of contaminating human plasma proteins, such as, particularly, the hepatitis virus.
• the stability of immunoglobulin in the heat treatment of this invention can be further enhanced by conducting the treatment in an inert gas atmosphere.
• the inert gas include nitrogen, argon and helium.
• the stabilizer used in this invention is at least one member selected from glycine, an alkalimetal chloride such as sodium chloride, an alkalimetal acetate such as sodium acetate, polyethylene glycol, albumin and mannitol.
• the stabilizer is used, for example, in an amount corresponding to the concentration thereof of about 0.01 to 4% (w/v) relative to 0.01 to 2% (w/v) immunoglobulin solution in the case of monoclonal immunoglobulin and relative to 2 to 8% (w/v) immunoglobulin solution in the case of polyclonal immunoglobulin.
• the stabilizing effect, the solubility in water and the state of solution are most well balanced for making the immunoglobulin into a medical preparation when the amount of the stabilizer added is in the above-mentioned range.
• % (w/v) means herein the amount of a solute by weight (gram) in 100 ml of the resulting solution.
• the immunoglobulin is usually used in a substantially dried, lyophilized state and the stabilizer is preferably added to an immunoglobulin solution prior to the lyophilization treatment thereof.
• the stabilizer may be removed after the dry heat treatment of this invention from the immunoglobulin preparation, it may be left incorporate therein as far as permitted.
• the heat treatment of this invention is preferably carried out with respect to a fraction or a composition containing purified immunoglobulin so as to be administered to a patient, it may be conducted at any step of the purification of the immunoglobulin.
• the process of this invention enables the inactivation of viruses, which may possibly contaminate an immunoglobulin preparation, without losing much of the activity of immunoglobulin, a precious plasma protein, it is useful as an industrial process for producing the plasma protein preparation.
• Fr-II (IgG fraction) was obtained from normal human plasma by means of the Cohn's cold alcohol fractionation method.
• One kg of the Fr-II paste thus obtained was dissolved in 1.5 l of cold water and then mixed with 15 g of glycine.
• the resulting IgG solution (5% w/v) was adjusted to a pH of 6.3 to 6.5 and then lyophilized. The moisture content after the lyophilization was 0.8%.
• the lyophilized IgG powder was heat-treated at 60° C. for 72 hours.
• the product thus obtained was examined for test items of solubility, HBsAg antibody titer, measles antibody titer, anti-diphtherial toxin titer, electrophoresis using a cellulose acetate membrane, and gel filtration in comparison with the IgG before the heat treatment. The results have revealed that no marked change is observed in any of these items and human IgG is stable under the above-mentioned heat treatment conditions.
• Fr-III fraction obtained from normal human plasma by the Cohn's cold alcohol fractionation method was purified by the method of salting-out or fractionation using acrinol.
• a 5% (w/v) solution of the human IgA thus obtained was mixed with glycine to its final concentration of 1% (w/v), and sodium chloride to that of 0.5% (w/v), and the mixed solution was lyophilized.
• the moisture content of the lyophilized powder was 2% or less.
• the powder was heated at 60° C. for 72 hours and then examined for its solubility, IgA concentration, measles antibody titer, and electrophoresis, using a cellulose acetate membrane. The test results were compared with those obtained with the powder before the heating and revealed that the powder was stable also after the heat treatment.
• the antibody titer against human lymphoblasts and the solubility of the lyophilized products were tested both before and after the heat treatment. The results have revealed that 75% of the antibody titer remains and the solubility shows no change after the above-mentioned heat treatment.
• mice monoclonal antibodies against established human cancer cells (MKN-45) belonging to the IgG class
• sodium chloride to a final concentration of 0.6% (w/v) and mannitol to that of 1% (w/v)
• the resulting solution was lyophilized to a substantially dry state.
• the lyophilized product was heated at 60° C. for 72 hours. The solubility and the antibody titer were tested with the lyophilized product before and after the heat treatment and the results have revealed that they are not changed by the above-mentioned heat treatment.
• the results have revealed that the stability is improved by the use of the stabilizer as compared with the case where no stabilizer is added (control).
• the stabilizing effect was similar both in the single use of a stabilizer and in the joint use of two or more stabilizers. Further, no particular difference in the effect was observed with difference in the mixing ratio of jointly used stabilizers.
• the effective amount of the stabilizer is 0.01 to 2% (w/v) relative to the aqueous solution of the IgG fraction.
• the effective amount was the same irrespective of whether the stabilizers were used each alone or in combination.

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• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Medicinal Chemistry (AREA)
• Chemical & Material Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• Animal Behavior & Ethology (AREA)
• Epidemiology (AREA)
• Pharmacology & Pharmacy (AREA)
• Mycology (AREA)
• Microbiology (AREA)
• Engineering & Computer Science (AREA)
• Immunology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
• Preparation Of Compounds By Using Micro-Organisms (AREA)
• Peptides Or Proteins (AREA)
• Micro-Organisms Or Cultivation Processes Thereof (AREA)

Applications Claiming Priority (2)

Publications (1)

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Cited By (24)

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Citations (9)

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• 1984
  • 1984-09-25 JP JP59200241A patent/JPH0669961B2/ja not_active Expired - Lifetime
• 1985
  • 1985-09-19 CA CA000491110A patent/CA1268707A/en not_active Expired - Lifetime
  • 1985-09-23 ES ES547212A patent/ES8700689A1/es not_active Expired
  • 1985-09-23 US US06/778,708 patent/US4721777A/en not_active Expired - Fee Related
  • 1985-09-24 KR KR8507004A patent/KR920005690B1/ko not_active Expired
  • 1985-09-24 EP EP85112134A patent/EP0177836B1/de not_active Expired - Lifetime
  • 1985-09-24 DE DE8585112134T patent/DE3577802D1/de not_active Expired - Lifetime

Patent Citations (10)

Non-Patent Citations (2)

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